Study of antifungal agent caspofungin adsorption to laboratory materials.

Treatment of invasive fungal infections with Caspofungin is used as the first-line antifungal agents. The minimum inhibitory concentration value is a test which indicates the degree of sensitivity of a strain regarding a drug. However, no value of minimum inhibitory concentration for caspofungin is available because very variable value is obtained. In this work, we study the link with the adsorption phenomenon of CSF previously described in literature and the lack of minimum inhibitory concentration value. A systematic study of the impact of different parameters on CSF adsorption is reported. The effect of the nature of container material, the aqueous solution pH and the organic solvent proportion was studied. In addition, the possibility of using a coating agent to minimize the adsorption was assayed and evaluated. Results obtained showed the importance of the material used during the manipulation of CSF. The use of acidic pH aqueous solution or the addition of acetonitrile or methanol proportions (50 % and 70 %, respectively) were found efficient to avoid adsorption of CSF on glassware material, which is the relevant strategy for analytical samples of caspofungin. The treatment of HPLC glass vials and 96-well plates with N-(2-aminoethyl)-3-aminopropyltrimethoxysilane reduced the adsorption. The significant adsorption observed in this work especially with plastic materials, questions the results obtained before in different assays and explained the absence of MIC value.

Estimation of the post-mortem interval of human skeletal remains using Raman spectroscopy and chemometrics.

An important demand exists in the field of forensic analysis to objectively determine the post-mortem interval (PMI) when human skeletal remains are discovered. It is widely known that bones undergo different chemical and physical processes after death, mainly due to their interaction with the environment in which they are found, although it is not known exactly what these processes consist of. Multiple techniques have been used so far to follow up these and other post-mortem changes and thus establish the time elapsed since the individual's death, but they present important drawbacks in terms of reliability and accuracy. The aim of this research was to propose an analytical methodology capable of determining the PMI by using non-destructive Raman spectroscopy measurements of human skeletal remains. The recorded Raman spectra provided valuable and potentially useful information from which a multivariate study was performed by means of orthogonal partial least squares regression (OPLSR) in order to correlate the PMI with the detected spectral modifications. A collection of 53 real human skeletal remains with known PMI (15 years ≤ PMI ≤ 87 years) was analysed and used for building and validating the OPLS model. The PMI of 10 out of 14 validation samples could be determined with an accuracy error of less than 30%, demonstrating the adequate predictive performance of the OPLS model even in spite of the large inter-individual variability it handled. This opens up the possibility of applying the OPLS model in combination with non-destructive techniques to the determination of the PMI of human skeletal remains that have been buried in conditions similar or equal to those of cemetery niches and in a geographic location with a Mediterranean climate, which is an important achievement for forensic medicine and anthropology.

Determination of antifungal caspofungin in RPMI-1640 cell culture medium by column-switching HPLC-FLD.

The actual scenario in the fight against fungal infections forces researchers to carry through with resistance studies to improve the therapies. These studies, which are performed in cell culture media, need accurate and sensitive analytical methodologies. That is why, in this work, an analytical method for caspofungin (CSF) concentration determination in RPMI-1640 cell culture medium with on-line sample treatment was developed and validated. CSF concentration was determined by HPLC-FLD using a column-switching procedure. The chromatographic analysis was carried out in less than 10 min using a C8 column (4 × 4 mm, 5 µm) as extraction stationary phase and a HSS T3 column (4.6 × 100 mm, 5 µm) as the analytical column. The used mobile phases were mixtures of phase A: pH 2 (adjusted with TFA) aqueous phase and phase B: ACN. For the extraction, the composition was (95:5, A:B v/v) and for the analysis (60:40, A:B v/v), both done in isocratic elution mode. These chromatographic conditions allowed reaching a limit of quantification of 10 µg/L, using 100 µL of sample with an injected volume of 40 µL. The proposed method was successfully validated in terms of selectivity, carryover, linear concentration range, accuracy and precision according to the criteria established by the Food and Drug Administration. Available amount of CSF in RPMI-1640 solution was found critical. CSF concentrations remained stable up to 2 h at room temperature. The developed method was applied for the direct analysis of CSF concentrations from in vitro experiments in presence of C. glabrata (CAGL18). The results highlight the decrease of cell proliferation even if the CSF amount decreases too, which asks question about the real value of the efficient concentration for CSF antifungal activity.

Successful Outcome Following Intravenous Lipid Emulsion Rescue Therapy in a Patient with Cardiac Arrest Due to Amitriptyline Overdose.

BACKGROUND The management of patients with tricyclic antidepressant drug overdose can be a challenge for the emergency department physician. Tricyclic antidepressants block alpha-adrenergic receptors and the anticholinergic effects may lead to cardiotoxicity, resulting in arrhythmias and hypotension that can lead to patient mortality. This report is of a case of a 28-year-old woman who presented with cardiac arrest due to amitriptyline overdose and who responded to intravenous lipid emulsion (ILE) therapy. CASE REPORT A 28-year-old woman was admitted to the emergency department with amitriptyline overdose. She suffered a cardiac arrest followed by cardiovascular and neurological complications. Hypotension and lack of a pulse did not respond to treatment with high-dose sodium, but she stabilized following treatment with ILE. The prompt response from the emergency team guaranteed rapid intervention that may have influenced the successful results. CONCLUSIONS Despite the frequency and severity of poisoning with tricyclic antidepressants, there is little consensus among physicians regarding patient management. This case showed the successful use of ILE as rescue therapy in a patient in cardiac arrest following amitriptyline overdose. However, the successful outcome obtained in this case is not a recommendation for the use of ILE as a first-line treatment for the management of patients with tricyclic antidepressant drug overdose. Controlled clinical studies are required to evaluate the safety and efficacy of ILE in the management of tricyclic antidepressant drug overdose.

Cleaning efficiency enhancement by ultrasounds for membranes used in dairy industries.

Membrane cleaning is a key point for the implementation of membrane technologies in the dairy industry for proteins concentration. In this study, four ultrafiltration (UF) membranes with different molecular weight cut-offs (MWCOs) (5, 15, 30 and 50kDa) and materials (polyethersulfone and ceramics) were fouled with three different whey model solutions: bovine serum albumin (BSA), BSA plus CaCl2 and whey protein concentrate solution (Renylat 45). The purpose of the study was to evaluate the effect of ultrasounds (US) on the membrane cleaning efficiency. The influence of ultrasonic frequency and the US application modes (submerging the membrane module inside the US bath or applying US to the cleaning solution) were also evaluated. The experiments were performed in a laboratory plant which included the US equipment and the possibility of using two membrane modules (flat sheet and tubular). The fouling solution that caused the highest fouling degree for all the membranes was Renylat 45. Results demonstrated that membrane cleaning with US was effective and this effectiveness increased at lower frequencies. Although no significant differences were observed between the two different US applications modes tested, slightly higher cleaning efficiencies values placing the membrane module at the bottom of the tank were achieved.

cBid, Bax and Bcl-xL exhibit opposite membrane remodeling activities.

The proteins of the Bcl-2 family have a crucial role in mitochondrial outer membrane permeabilization during apoptosis and in the regulation of mitochondrial dynamics. Current models consider that Bax forms toroidal pores at mitochondria that are responsible for the release of cytochrome c, whereas Bcl-xL inhibits pore formation. However, how Bcl-2 proteins regulate mitochondrial fission and fusion remains poorly understood. By using a systematic analysis at the single vesicle level, we found that cBid, Bax and Bcl-xL are able to remodel membranes in different ways. cBid and Bax induced a reduction in vesicle size likely related to membrane tethering, budding and fission, besides membrane permeabilization. Moreover, they are preferentially located at highly curved membranes. In contrast, Bcl-xL not only counterbalanced pore formation but also membrane budding and fission. Our findings support a mechanism of action by which cBid and Bax induce or stabilize highly curved membranes including non-lamellar structures. This molecular activity reduces the energy for membrane remodeling, which is a necessary step in toroidal pore formation, as well as membrane fission and fusion, and provides a common mechanism that links the two main functions of Bcl-2 proteins.

Oscillations in the evaluation of fractal dimension of RR intervals time series.

Previously, we have reported the presence of oscillations in the graphs we have used to evaluate the Higuchi's fractal dimension in RR intervals time series of congestive heart failure (CHF) patients in the sleep phase but these oscillations hardly appear in all the six hours of the awake phase. In this paper we report the same analysis for heart rate time series for different groups of healthy subjects; we are looking for the presence of this kind of oscillations in other situations. We analyzed all the time series in the Exaggerated Heart Rate Oscillations database of Physionet during two meditation techniques: volunteers with spontaneous breathing, subjects in meditation, volunteers in a metronomic breathing group and elite athletes. We have found oscillations in the graphs of the Higuchi's fractal dimension in the heart rate time series of subjects in meditation and metronomic breathing and this fact coincides with previous reported results.

Acute effects of passive stretching and vibration on the electromechanical delay and musculotendinous stiffness of the plantar flexors.

The purpose of the present study was to examine the acute effects of passives stretching versus prolonged vibration on the active and passive properties of voluntary and evoked muscle actions of the plantar flexors. Eleven healthy men performed the isometric maximal voluntary contractions (MVCs) and passive range of motion (PROM) assessments before and after 20 min of passive stretching (PS), vibration (VIB), and control (CON) conditions. In addition, percent voluntary activation was calculated from superimposed and potentiated doublets during the MVCs. Voluntary peak torque (PT) decreased by 11% and 4%, while surface electromyographic (EMG) amplitude decreased by 8% and 16% for the PS and VIB, respectively, with no changes during the CON The electromechanical delay (EMD) decreased and PROM increased following the PS, but was unchanged during the VIB and CON conditions. Musculotendinous stiffness (MTS) decreased at all joint angles following the PS, but decreased only at the furthest joint angle following the VIB. There were no changes in peak twitch torque (PTT), M-wave amplitude, and EMG amplitude during the PROM assessments for all conditions. Both PS and VIB elicited similar decreases in muscle activation, which may be the same centrally-mediated mechanism (i.e., y loop impairment). Changes in the EMD were inversely proportional to the changes in MTS, which occurred only following PS. The present findings indicated that the stretching- and vibration-induced force deficits may have resulted in part from similar centrally-mediated neural deficits, while an elongation of the series elastic component may also have affected the stretching-induced force deficit.

Correlation of the genotoxic activation and kinetic properties of Salmonella enterica serovar Typhimurium nitroreductases SnrA and cnr with the redox potentials of nitroaromatic compounds and quinones.

Bacterial nitroreductases (NRs) catalyse the oxygen-insensitive reduction of several nitro-substituted compounds and quinones. SnrA and cnr NRs have been previously identified in Salmonella enterica serovar Typhimurium; they reduce several environmental nitro compounds that display mutagenic activity in the Ames test. Although some of their biochemical properties have been reported, the substrate specificity of each protein over mutagenic nitro compounds is unknown; even more, the possible relationship between their capacity to activate nitro compounds into mutagens and the redox properties of putative substrates has been poorly investigated. We have purified SnrA and cnr and investigated their capacity to activate several mutagens in the Ames test as well as their kinetic parameters K(m) and V(max). Our results show that SnrA and cnr are able to activate 2,7-dinitrofluorene with the same efficiency and a similar mutagenic potency in the YG7132 tester strain; 1-nitropyrene and 1,3-dinitropyrene were efficiently activated by cnr, whereas 1,8-dinitropyrene, 1,6-dinitropyrene and 2-nitrofluorene were scarcely activated by either NR. The mutagenic potency of nitro compounds obtained in the presence of either enzyme correlates with their redox potential reported in the literature. On the other hand, a good correlation was obtained between the catalytic efficiency (V(max)/K(m)) of the purified cnr with the redox potential of eight molecules including nitro-substituted compounds and quinones. No correlation between redox potential and catalytic efficiency by SnrA was observed, suggesting that factors other than redox potential such as the structure of the compounds are involved in the catalytic efficiency of SnrA.

Analysis of RR intervals time series of congestive heart failure patients with Higuchi's fractal dimension.

We study time series of RR intervals of subjects with normal sinus rhythm (NSR) during sleep and wake phases and we also analyze time series of subjects with congestive heart failure (CHF) in both phases with the method of Higuchi's fractal dimension. We have found the presence of oscillations in the plots we have used to evaluate the Higuchi's fractal dimension; these oscillations seem to be associated with the appearance of periodicities in the time series of CHF patients in the sleep phase. These periodicities do not appear in all the six hours of the sleep phase, so this regular behavior is not observed in all sleep stages. It could be possible that patients that always show these periodicities are in worst condition, and it is possible that the disease worsens when periodicities appear in the Higuchi's graph.

Comparison of the behavior of two nanofiltration membranes for sweet whey demineralization.

Nanofiltration is a process used to separate mineral salts from lactose, having previously removed the proteins by ultrafiltration. Both proteins and lactose can be used as raw materials to prepare a variety of products. In this paper, we studied the feasibility of demineralizing sweet whey obtained from the cheese industry of the Comunidad Valenciana (Spain) using membrane technologies. The NF200 membrane showed the highest volumetric flux and solute rejection values, whereas the DS-5 DL membrane showed the lowest values. The volumetric fluxes obtained with the NF200 and DS-5 DL membranes in these experiments with the ultra-filtered whey demonstrated significant differences between membranes. Concerning solute rejection, the highest values were obtained using the NF200 membrane. The chosen parameter to evaluate the demineralization capability was solute flux. In this way, the values obtained for chloride ion were 9.90 and 32.42 g/(m2 x h) for the NF200 and DS-5 DL membranes, respectively, with the highest demineralization rates being achieved with the DS-5 DL membrane.

Electro-Fenton as a feasible advanced treatment process to produce reclaimed water.

The feasibility of the electro-Fenton process to generate simultaneously both of the Fenton's reagent species (Fe2+/H2O2), was assessed as a potentially more economical alternative to the classical Fenton's reaction to produce reclaimed water. An air-saturated combined wastewater (mixture of municipal and laboratory effluents) was treated in discontinuous and continuous reactors at pH = 3.5. The discontinuous reactor was a 2 L electrochemical laboratory cell fitted with concentric graphite and iron electrodes. The continuous reactor tests used a pilot treatment system comprising the aforementioned electrochemical cell, two clarifiers and one sand filter. Several tests were carried out at different conditions of reaction time (0-60 min) and electrical current values (0.2-1.0 A) in the discontinuous reactor. The best operating conditions were 60 min and 1 A without filtration of effluents. At these conditions, in discontinuous and continuous reactors with filtration, the COD, turbidity and color removal were 65-74.8%, 77-92.3% and 80-100%, respectively. Fecal and total coliforms, Escherichia coli, Shigella and Salmonella sp. were not detected at the end of the pilot treatment system. Electrogeneration of the Fenton's reagent is also economical; its cost is one-fifth the cost reported for Advanced Primary Treatment.

The influence of an internal electric field upon protein crystallization using the gel-acupuncture method.

In this work, the influence of an internal electric field upon the crystallization of lysozyme and thaumatin is explored using a modified design of the gel-acupuncture setup. From a crystallographic point of view, the orientation of crystals that grow preferentially over different types of electrodes inside capillary tubes is also evaluated. Finally, the crystal quality and the three-dimensional structure of these proteins grown with and without the electric field influence are analyzed by means of X-ray diffraction methods.

Cloning and characterization of a FAD-monooxygenase gene ( cadA) involved in degradation of chloranilic acid (2,5-dichloro-3,6-dihydroxybenzo-1,4-quinone) in Pseudomonas putida TQ07.

A bacterium culture was isolated on the basis of its ability to degrade chloranilic acid, and was later identified as Pseudomonas putida (TQ07). Several transposon insertion mutants unable to degrade chloranilic acid were selected. The characterization of the site of insertion of one of these mutants led to the identification of the cadA gene encoding an enzyme with significant homology with FAD-monooxygenases involved in the degradation of aromatic and chloroaromatic compounds. The finding that, after replacing the mutant allele with the wild-type one, the strain recovered the wild-type pattern of "halo" formation (a zone of clearing color on agar plates around TQ07 colonies that degrade chloranilic acid) and degradation of chloranilic acid, unequivocally assigned cadA a function in the metabolism of this compound. We also found that most of the transposon insertion mutants unable to degrade chloranilic acid are clustered in a 10-kb region of the P. putidagenome that is encoded in a megaplasmid or in an unstable chromosomal region.

Nemorosone, the major constituent of floral resins of Clusia rosea.

Nemorosone, the major constituent of the floral resin of Clusia rosea was isolated after exhaustive chromatography. This compound was fully characterized as it is in the nature, without methylation as reported before. A keto-enol equilibrium was observed and both isomers were totally characterized by NMR spectroscopic techniques. The previously announced structure for methylnemorosone was corrected on the basis of application of chemical methylation, high field 2D NMR techniques and NOE difference spectroscopy experiments on the natural product. Our studies concluded that an interchange occurred in the assignment of the benzoyl moiety position with an isoprenyl group in that structure.

Prevalencia de la esclerosis múltiple en Colombia / Prevalence of multiple sclerosis in Colombia

Introducción. La esclerosis múltiple (EM) es la enfermedad desmielinizante más común del sistema nervioso. Se caracteriza por numerosas áreas desmielinizantes o placas de desmielinización que se encuentran dispersas en el sistema nervioso. Se ha demostrado que la EM es menos frecuente en las regiones tropicales que en aquellas localizadas en el subtrópico, En Latinoamérica, particularmente, existen algunos estudios que evidencian dicho fenómeno.Sin embargo, en Colombia no se han realizado estudios de prevalencia de EM. Objetivo. Determinar la prevalencia de la esclerosis múltiple en cinco provincias colombianas (Antioquia, Caldas, Santander, Risaralda y Bolívar). Pacientes y métodos. Se utilizó el método de captura-recaptura de dos fuentes para estimar el número de casos de EM definida según los criterios de Poser et al, durante el período comprendido entre julio de 1995 y junio de 2000. Resultados. La prevalencia (casos de EM por 100.000 habitantes) varió entre 1,48 en Antioquia (IC95 por ciento 1,12; 1,78) a 4,98 en Risaralda (IC95 por ciento 3,52; 6,43). El 72 por ciento de los individuos identificados eran mujeres. Las regiones incluidas en este estudio comprenden al 25 por ciento de la población de Colombia. Conclusiones. La prevalencia de EM estimada es baja y está de acuerdo a lo esperado para las áreas tropicales. Los individuos con EM de estas regiones pueden ser muy útiles para el estudio de otros factores involucrados en la etiología de la EM (genético). El método de captura- recaptura es una excelente herramienta para la realización de estudios de prevalencia porque consume poco tiempo y es muy barato (AU)

[Prevalence of multiple sclerosis in Colombia]. / Prevalencia de la esclerosis múltiple en Colombia.

INTRODUCTION: Multiple sclerosis (MS) is the commonest demyelinating condition of the nervous system. It is characterized by numerous demyelinating areas or plaques of demyelination which are found dispersed throughout the nervous system. It has been shown that MS is less frequent in tropical regions than in subtropical regions. In Latin America particularly, there are some studies which show this phenomenon. However, in Colombia no studies of prevalence of MS have been done. OBJECTIVE: To determine the prevalence of multiple sclerosis in five provinces of Colombia (Antioquia, Caldas, Santander, Risaralda and Bolivar). PATIENTS AND METHODS: The capture-recapture method was used for two sources to determine the number of cases defined on the criteria of Poser et al seen between July 1995 and June 2000. RESULTS: The prevalence (cases of MS per 100,000 inhabitants) varied between 1.48 in Antioquia (95% CI 1.12; 1.78) and 4.98 in Risaralda (95% CI 3.52; 6.43). Seventy two percent were women. The regions included in this study represented 25% of the population of Colombia. CONCLUSIONS: There is a low prevalence of MS which is as expected in tropical areas. Persons with MS in these regions may be very useful in the study of other factors involved in the aetiology of MS (genetic). The capture-recapture method is an excellent tool for carrying out prevalence studies since it is cheap and requires little time.

Estudio de relación padres/hijos en adolecentes escolares en la Unión / Study of parent-child relation in school adolescents in la Unión

Se encuestaron 385 estudiantes de ambos sexos del 3§ y 4§ medio del Liceo B-12 de la Unión y a 98 de sus padres, encontrándose que un 72,2% vive con ambos padres. La mitad de los hijos tiene más confianza con su madre y un tercio con ninguno. El 51,9% de los hijos afirma haber recibido educación sexual de sus padres. La mayoría de los hijos y sus padres considera que la calidad del tiempo dedicado a ellos es más importante que la cantidad. Un 13% de los hijos no sabe si sus padres les quieren. Los conflictos padres/hijos variaron según el sexo. Las principales demandas de los hijos fueron más libertad, más cariño y más respeto. El 25,8% de las hijas y 40,0% de los hijos manifestó estar conforme con sus padres. Se establece la necesidad de mejorar la comunicación y acercamiento afectivo entre padres e hijos

Asociacion entre campylobacter pylori y gastritis: metodos de identificacion: estudio de 99 casos en el Hospital San Ignacio / Association between campylobacter pyloridis and gastritis: identification method: study of 99 cases in Hospital San Ignacio

Se estudian 99 biopsias gastricas incluyendo casos de gastritis y controles normales, identificando el campylobacter pylori por los metodos de coloracion en tejido, prueba de la ureasa y cultivo. Se correlaciona el tipo de gastritis histologicamente con la presencia o no de la bacteria y se analiza la utilidad de cada uno de los metodos de identificacion